Fig. (2).

Codon usage bias adjustment of optimized gene for expression in E.coli (A); The percentage distribution of codons in computed codon quality groups. The value of 100 is set for the codon with the highest usage frequency for a given amino acid in the desired expression organism. Codons with values lower than 30 are likely to hamper the expression efficiency (B); The distribution of codon usage frequency along the gene sequence. A CAI of 1.0 is measured to be perfect in the desired expression organism, and a CAI of >0.8 is considered as good, in terms of high gene expression levels (C); GC content analysis of optimized chimeric gene; the ideal percentage range of GC content is between 30% to 70%. Any peaks outside of this range will adversely affect transcriptional and translational efficiency.